Study on Skin Care Efficacy of the Active Ingredients in Camellia Japonica Flowers at High and Low Altitudes

The active ingredients of Camellia japonica flowers(CJF)at high and low altitudes,as well as their skin care efficacy were compared.The dried red CJF at high and low altitudes were ultrasonically extracted with 70%ethanol,and these extracts were concentrated and then diluted to a constant volume.The content of total flavonoids,total polyphenols and total proteins was tested and analyzed.In addition,DPPH free radical scavenging,inhibition of the formation of advanced glycation end products,and inhibitory activity against elastase was measured to compare their skin care efficacy in anti-oxidation,anti-glycation,anti-wrinkling and firming.The results showed that,based on the dry weight of CJF,the content of total flavonoids and total proteins of the CJF at high altitude was approximately 1.5 times of the CJF at low altitude,while the content of total polyphenols was approximately 2.4 times that of the CJF at low altitude.The skin care efficacy of CJF at high altitude was significantly better than that of the CJF at low altitude.This work could provide theoretical basis for the selection and application of Camellia japonica flowers in the field of cosmetics.

Collecting Knowledge about Medicinal Ingredients in Northwestern Sichuan in the 1950s

This article explores China’s state-level knowledge production on medicinal ingredients in northwestern Sichuan in the 1950s.Drawing on county-level archives,published materials and interviews,this historical article traces how different levels of governments and state-owned trading companies produced knowledge about medicinal ingredients and its production.It argues that on one hand the state’s procurement standards codified the knowledge about medicinal ingredients from the marketplace and the local producers.On the other hand,direct extraction of knowledge from the medicine gatherers and cultivators verbalized and collected the previously tacit local knowledge about the production of medicinal ingredients,which would help to alleviate the shortage in the supply of traditional Chinese medicines throughout the 1950s.

A review on cocrystal of active ingredients in traditional Chinese medicine

In the development of new drugs products,especially the development of traditional Chinese medicine active ingredients,solubility and oral bioavailability are the main factors which are restricting the development of new drugs,whereas the physicochemical properties of active ingredients are the key element to affecting these factors.Pharmaceutical cocrystal provides an excellent opportunity to develop new drugs with excellent physical and chemical properties such as melting point,solubility,stability and bioavailability while retaining the pharmacological properties of individuals active pharmaceutical ingredients among pharmaceutical cocrystal compounds.Traditional Chinese herbal medicine has the characteristics of multiple pathways and multiple targets,mainly because it contains many active ingredients,like cocrystals thereof with many components.The active ingredients extracted from traditional Chinese herbal medicine have a wide range of pharmacological activities,but most of the active ingredients affect the development of traditional Chinese medicine active ingredients due to their physical and chemical properties such as solubility.Traditional Chinese medicine pharmaceutical cocrystals can not only improve the physical and chemical properties of drugs without changing the internal structure of drugs,so as to provide a new scheme for the development of traditional Chinese medicine active ingredients.This paper reviews the research progress of active ingredients of traditional Chinese medicine pharmaceutical cocrystal.The preparation methods of cocrystals are summarized and the advantages of cocrystals are illustrated with examples.

Application of Caenorhabditis elegans in screening active ingredients of traditional Chinese medicine

People continue to recognize the important role of Chinese medicine in the treatment of diseases,and there is an urgent need to clarify the mechanism of Chinese medicine.Because of the multi-target and multi-component characteristics of traditional Chinese medicine,it is difficult to elaborate from a single perspective.As a high-quality animal model,Caenorhabditis elegans with the characteristics of small size and fast reproduction,has been widely used in traditional Chinese medicine activity experiments.In this article,Caenorhabditis elegans,as model organisms used to screen the active components of traditional Chinese medicine were reviewed.

Methodologies on estimating the energy requirements for maintenance and determining the net energy contents of feed ingredients in swine:a review of recent work

In the past two decades, a considerable amount of research has focused on the determination of the digestible(DE) and metabolizable energy(ME) contents of feed ingredients fed to swine. Compared with the DE and ME systems, the net energy(NE) system is assumed to be the most accurate estimate of the energy actually available to the animal. However, published data pertaining to the measured NE content of ingredients fed to growing pigs are limited. Therefore, the Feed Data Group at the Ministry of Agricultural Feed Industry Centre(MAFIC) located at China Agricultural University has evaluated the NE content of many ingredients using indirect calorimetry. The present review summarizes the NE research works conducted at MAFIC and compares these results with those from other research groups on methodological aspect. These research projects mainly focus on estimating the energy requirements for maintenance and its impact on the determination, prediction, and validation of the NE content of several ingredients fed to swine. The estimation of maintenance energy is affected by methodology, growth stage,and previous feeding level. The fasting heat production method and the curvilinear regression method were used in MAFIC to estimate the NE requirement for maintenance. The NE contents of different feedstuffs were determined using indirect calorimetry through standard experimental procedure in MAFIC. Previously generated NE equations can also be used to predict NE in situations where calorimeters are not available. Although popular, the caloric efficiency is not a generally accepted method to validate the energy content of individual feedstuffs. In the future,more accurate and dynamic NE prediction equations aiming at specific ingredients should be established, and more practical validation approaches need to be developed.

Effects of dietary fiber content and different fiber-rich ingredients on endogenous loss of fat and fatty acids in growing pigs

Background: Determination of the endogenous loss of fat(ELF) is used to adjust for the estimation of true total tract digestibility(TTTD) of fat in diets and ingredients.Any factor which affected ELF may further affect the digestibility of fat,including sources and concentrations of fat and fiber in the diet.There are some reports of determining the ELF using regression methods based on different levels of fat intake,while reports on effects of dietary fiber content and different fiber-rich ingredients in pig diets on ELF are very limited.Therefore,the objective of this study was to determine the effects of dietary fiber content and different fiber-rich ingredients on endogenous losses of fat and fatty acids at the end of ileum and throughout the entire intestinal tract in growing pigs.Methods: In Exp.1,the effect of fiber content on endogenous loss of fat was determined using six growing pigs(Duroc × Landrace × Yorkshire;27.6 ± 2.4 kg),fitted with a T-cannula at the end of ileum.The experimental design was a 6 × 6 complete Latin square design with six periods of feeding and six diets.The six experimental fat-free diets were formulated to include graded levels of neutral detergent fiber(NDF)(0,40,80,120,160 and 200 g/kg) and soybean hull(SH) was the only fiber source,providing 0,75,150,225,300 and 375 g/kg,respectively.Chromic oxide was included at4 g/kg in all diets as an indigestible marker.In Exp.2,six crossbred growing barrows(27.6 ± 1.6 kg) were used and the experimental design was the same as for Exp.1.The six fat-free diets were formulated to include six common fiber-rich ingredients and the concentration of NDF was 100 g/kg.The six fiber-rich ingredients were defatted rice bran(DRB),sugar beet pulp(SBP),rice hull(RH),corn germ meal(CGM),SH and wheat bran(WB) and they were fed at represented250,270,145,250,170 and 280 g/kg in the diet,respectively.Results: In Exp.1,the endogenous loss of fatty acids profile did not change as dietary NDF increased in growing pigs.The endogenous losses of fat,C16:0,C18:0,C18:1,C18:2,total unsaturated fatty acids(UFA) and total saturated fatty acids(SFA) in growing pigs at the end of ileum and throughout the entire intestinal tract increased linearly as NDF content of diets increased.The endogenous losses of fat,as well as C16:0 and C18:0 throughout the entire intestinal tract also increased quadratically as NDF content of diets increased.The ELF increased from 0.71 to 3.14 g/kg of dry matter intake(DMI) and 0.56 to 8.21 g/kg DMI at the end of ileum and throughout the entire intestinal tract in growing pigs,respectively.The ELF occurred in the hindgut except for the growing pigs fed 0 and 4% NDF in their diets.The endogenous losses of C16:0 and UFA occurred primarily in the upper regions of the gut and the greatest endogenous losses of C18:0 occurred in the hindgut.The endogenous losses of fat,individual SFA and total SFA throughout the entire intestinal tract were much greater than that at the end of ileum.However,the endogenous losses of individual UFA and total UFA were less throughout the intestinal tract than at the end of ileum.In Exp.2,the endogenous losses of fat at the end of ileum were greater in growing pigs fed CGM or WB diets.The endogenous loss of fatty acids profile changed to a slight degree at the end of ileum that the endogenous loss of UFA(particularly C18:1 and C18:2) in growing pigs fed CGM or WB diets were greater(P < 0.01) than that for the other four diets.The greatest(P < 0.01) endogenous loss of SFA(particularly C18:0) was in growing pigs fed the RH diet.The endogenous losses of fat,C16:0,C18:0 and SFA over the entire intestinal tract were much greater in growing pigs fed CGM or WB diets,whereas the lowest values were in growing pigs fed DRB diet.The ELF at the end of ileum in growing pigs fed CGM or WB diets were 3.50 or 4.17 g/kg DMI,respectively,and the ELF over the entire intestinal tract was 7.23 or 7.38 g/kg DMI.The contribution in percentage of ELF in the upper gut was greater than that in the hindgut of growing pigs fed DRB and RH diets,while the ELF in the upper gut and hindgut were equal in growing pigs fed SBP,CGM and WB diets.On the whole,the endogenous losses of C18:1 and C18:2 throughout the entire intestinal tract in growing pigs fed the six fiber-rich ingredients diets were less than losses at the end of ileum,whereas the endogenous loss of fat,C16:0,C18:0 and SFA were greater throughout the intestinal tract than at the end of ileum.Conclusion: The profile of loss in endogenous fatty acids did not change as dietary NDF increased in growing pigs and the endogenous losses of fatty acids(C16:0,C18:0,C18:1 and C18:2) fat,UFA and SFA increased linearly as NDF content increased in the diets of pigs.The endogenous losses of fat or fatty acids at the end of ileum were greater in growing pigs fed RH,CGM or WB diets.The endogenous losses of fat,fatty acids(C16:0 and C18:0) and SFA were greater over the entire intestinal tract in pigs fed CGM or WB diet,while these values were the lowest in growing pigs fed the DRB diet.The contribution in percentage losses of fat in the upper gut were greater than in the hindgut of growing pigs fed DRB and RH diets,while the contribution of losses of fat in the upper gut and hindgut were equal in growing pigs fed SBP,CGM and WB diets.In addition,the endogenous loss of individual or total UFA was less over the entire intestinal tract of growing pigs fed fiber diets than that at the end of ileum,and the greatest endogenous losses of fat,individual or total SFA were over the entire intestinal tract.Therefore,differences in fiber content and the nature of fiber-rich ingredients in diets of pigs have different effects to the endogenous losses of fat or fatty acids.Considering the requirement of fat or fatty acids of pigs,careful attention must be paid that the endogenous losses of fat and fatty acids when fiber ingredients are used in diets of pigs.

Probiotical Cell Fragments (PCFs) as “Novel Nutraceutical Ingredients”

Probiotical cell fragments (PCFs) are structural components of the probiotic cell lysate(s) and exhibit similar beneficial effects on the host as live probiotic bacteria. With cell fragment technology (CFT?), the structural fragments are isolated and purified from live probiotic cells. While observed to be strain-dependent as in the case of live probiotics, orally administered PCFs demonstrated a broad spectrum of immune modulation functions;anti-allergy;anti-inflammation;anti-bacterial and anti-viral properties;anti-mutagenic;and radioprotective and detoxification abilities in humans and animals. The PCFs mechanisms of action include events of motifs of cell wall peptidoglycans, DNA motifs, nucleotide containing components, lipoteichoic acids (LPAs), surface layer (S-layer) proteins, and cytoplasmic proteins. Different immunological in vivo-in vitro tests have shown that PCFs, essentially, have the ability to stimulate the macrophages, and induce cytokines such as interleukins, tumor necrosis factors (TNFs), interferons (IFNs), and natural killer (NK) cells. PCFs may be used as ingredients for foods and beverages or as nutritional supplements with long term stability and shelf-life up to 5 years. PCFs may also be used as health restorative ingredients in cosmetic products. The outcome of probiotics CFT? stands as an advantage to the food and pharmaceutical industries, regarding the formulation of novel products with unadulterated sensory characteristics of origin. Hence, PCFs are being characterized here as “novel nutraceutical ingredients” for health maintenance in both humans and animals.

Structural similarity-based prediction of the potential active ingredients and mechanism of action of traditional Chinese medicine formulations used to anti-aging

Objective:To predict the potential active ingredients (PAIs) and mechanism of action of traditional Chinese medicine formulations used to delay aging.Methods:We incorporated the use of quantum-chemistry calculations and machine learning to predict the active ingredients of some Chinese herbal medicines used to delay aging.Then,a network-pharmacology approach was used to uncover how these PAls delayed aging.Results:Twelve PAls with anti-aging effects were discovered:androsterone,MHP,cortisone,propyl methyl trisulfide,retinol,retinal,cortisol,11-cis-Retinol (2R,3R)-3-hydroxyproline,4,5alpha-Dihydrocortisone (2S)-2-ammonio-6-ureidohexanoate and17alpha,21-Dihydroxy-5beta-pregnane-3,11,20-trione.Enrichment analyses indicated that a putative compound target and aging target were significantly associated with:regulation of the immune system;insulin receptor signaling pathway;regulation of the mitotic cell cycle;response to nutrient levels;response to oxidative stress;release of cytochrome c from mitochondria;learning or memory;inflammatory response.Conclusions:A novel method was proposed to predict the PAls of anti-aging herbal medicines by incorporating quantum-chemistry calculations and machine learning.Then,a network-pharmacology approach was used to uncover how these PAls delay aging.The information provided by our study on PAls may aid the discovery of anti-aging drugs.

Research progress on antidepressive active ingredients and mechanism of Xiaoyaosan

The antidepressive effect of Xiaoyaosan is doubtless by researches while the potential antidepressive ac- tive ingredients and their mechanisms remain unclear. In order to explain the antidepressive effect of Xiaoyaosan, all of the recent reports were sought out on the current situation of the antidepressive monomer compositions of Xi- aoyaosan and their mechanism. By exploring the relationship between the effects and the active ingredients, the mechanism and the active ingredients, the antidepressive active ingredients such as quercetin, isorhamnetin, isoliquiritigenin, saikosaponinA, saikosaponinD, glycyrrhiz- kaempferol, liquiritin, isoliquiritin, liquiritigenin, inate, paeoniflorin, albiflorin, ferulic acid, curcumin and their mechanism of antidepressive such as regulating the monoamine neurotransmitter systems, regulating the neuroendocrine system, affecting the neural plasticity and neu- affecting the cytokines level, anti-oxidative stress were rotrophic, affecting the cellular and molecular mechanisms , summarized. By the overview in the level of monomer rather than formulae, we uncovered the antidepressive mech- anism of Xiaoyaosan and further confirmed that Xiaoyaosan, as a classic antidepressive Chinese prescription, had many characteristics, such as multi-level , multi-channel, multi-target and so on .

Research Progress of Anti-tumor Active Ingredients in Dandelion

As a traditional Chinese herb,dandelion,containing complicated active ingredients which includes polysaccharide,flavonoid,terpene,pigment,phytosterol,coumarin,organic acid and etc,plays significant roles in various physiological activities in organisms.The active ingredients generally interfere in signal transduction of cancer cells,regulate cell cycle and apoptotic protein expression,inhibit cancer cells proliferation and migration and etc,which effectively restrains tumor development and deterioration.This article summarizes the anti-tumor mechanism of five active ingredients in dandelion through paper reading which provides thoughts and references for anti-tumor research.

Model evaluation for the prediction of solubility of active pharmaceutical ingredients(APIs) to guide solid–liquid separator design

The assumptions and models for solubility modelling or prediction in systems using nonpolar solvents, or water and complex triterpene and other active pharmaceutical ingredients as solutes aren't well studied. Furthermore, the assumptions concerning heat capacity effects(negligibility, experimental values or approximations) are explored, using non-polar solvents(benzene), or water as reference solvents, for systems with solute melting points in the range of 306–528 K and molecular weights in the range of 90–442 g/mol. New empirical estimation methods for the Δ_(fus)C_(pi) of APIs are presented which correlate the solute molecular masses and van der Waals surface areas with Δ_(fus)C_(pi). Separate empirical parameters were required for oxygenated and non-oxygenated solutes. Subsequently, the predictive capabilities of the various approaches to solubility modelling for complex pharmaceuticals,for which data is limited, are analysed. The solute selection is based on a principal component analysis, considering molecular weights, fusion temperatures, and solubilities in a non-polar solvent, alcohol, and water, where data was available. New NRTL-SAC parameters were determined for selected steroids, by regression. The original UNIFAC, modified UNIFAC(Dortmund), COSMO-RS(OL), and COSMO-SAC activity coefficient predictions are then conducted, based on the availability of group constants and sigma profiles. These are undertaken to assess the predictive capabilities of these models when each assumption concerning heat capacity is employed. The predictive qualities of the models are assessed, based on the mean square deviation and provide guidelines for model selection, and assumptions concerning phase equilibrium, when designing solid–liquid separators for the pharmaceutical industry on process simulation software. The most suitable assumption regarding Δ_(fus)C_(pi) was found to be system specific, with modified UNIFAC(Dortmund) performing well in benzene as a solvent system, while original UNIFAC performs better in aqueous systems.Original UNIFAC outperforms other predictive models tested in the triterpene/steroidal systems,with no significant influence from the assumptions regarding Δ_(fus)C_(pi).

Aflatoxin Contamination and Rancidity in Locally Processed Commercial Fish Feeds and Ingredients along the Value Chain in Wakiso and Kampala Districts, Uganda

Aflatoxin contamination and rancidity in locally processed commercial fish feeds and ingredients along value chains is a Public and Animal health hazard. The study established the level of aflatoxin contamination, peroxide value (PV), Anisidine value (AnV), and their associated factors at storage areas among farmers, processors, traders, factories, and landing sites in the Wakiso and Kampala districts. The value chain actors were purposively selected in a cross-sectional study based on access to the feed store and the use of locally processed commercial fish feeds on farms. Data collected were statistically analyzed in SPSS version 20. All the samples (45) were positive for aflatoxin contamination and PV with 51% (23/45) of samples being contaminated with “above acceptable” aflatoxin levels and 66.6% (30/45) of samples with “above acceptable” PV. The overall percentage of “above acceptable” AnV was 11/29 (37.9%). Samples from factories were within acceptable contamination levels. Multivariate logistic regression analysis revealed no significant difference between aflatoxin contamination, peroxide, and Anisidine value with storage factors for locally processed commercial fish feeds and ingredients. The study recommended the purchase of fish feeds from factories and a larger study on storage factors responsible for aflatoxin contamination and rancidity in fish feeds in Uganda.

Identification of Avian-Derived Ingredients in Livestock and Poultry Meat by PCR Technology

In order to establish a quick and specific method which could identify the avian-derived ingredients,this study used 16 S rRNA gene sequence as target site,and designed the specific primers of chicken,pigeon meat and quail meat. The DNA of common livestock and poultry meat( including mutton,beef,pork,rabbit meat,pigeon meat,quail meat,chicken,duck and goose) was used as template. Though PCR amplification and specific detection,a quick determination method was established to identify the avian-derived ingredients. The results showed that the selected primers could identify the ingredients of animal origin effectively and quickly. The method was convenient and concise,and could detect the chicken-derived,pigeon-derived,quail-derived ingredients in livestock and poultry food quickly and accurately.

Rapid screening of genetically modified ingredients in soybean and cotton processing by-product and waste using direct qPCR

To develop a simple and fast method for screening genetically modified ingredients from processing by-product and waste,direct quantitative PCR(qPCR)kit-Taqman which omitting multi genomic DNA preparing steps was developed in this study.A total of 18 oil crop processing by-products and wastes including 10 soybean and 8 cotton materials were collected from food processing factories.Compared with 2 commercial direct qPCR kits,conditions of DNA releasing procedure and PCR amplification were optimized.Element screening was performed at the initial step of genetically modified(GM)ingredient testing procedure via direct qPCR.GM event identification was carried out in positive samples by initial screening.Totally 5 screening elements(P–35S,T-NOS,Cp4-epsps,bar and pat)for soybean materials and 6 screening elements(P–35S,T-NOS,NPTII,Cry1Ac,bar and pat)for cotton samples were detected.In GM event identification,MON531 and MON1445 were found in cotton materials.Results were further confirmed by real-time PCR with DNA extraction and purification.The direct qPCR system proposed by this research was convenient for rapid screening and identification of GM ingredients in oil crop primary by-product and waste.

Multiple Emulsions Able to Be Used for Oral Administration of Active Pharmaceutical Ingredients: Physico-Chemical Parameters Study of Different Phases

Multiple emulsions are of great therapeutic interest especially in the administration of medicines which can be inactivated by digestive enzymes;moreover the researches of formulation not being often easy, a control of the different phases physicochemical parameters would be of great interest in rapid formulations and at low cost. When formulating emulsions, the preliminary tests, also known as formulation tests, constitute a step which can be long and expensive because of the quantity of reagents that can be used. A rigorous methodology could thus be of great interest, which is at the aim of our study which consists of evaluating the physico-chemical parameters of different phases used to make thus multiple emulsions. In our study, physico-chemical parameters such as conductivity, pH, density, viscosity, and surface tension have been studied by direct measurement using equipment and also by means of suitable mounting. The results showed that the pH and the surface tension have an important role in the prediction of the stability of emulsions, these latter must be of the same order of magnitude. For all phases conductivity does not have too much interest apart from helping to determine the type of the emulsion.

New development of whey protein products as nutritional food ingredients

Whey protein products provided ideal ingredients for nutritional food applications.Whey proteins are known to have high nutritional quality due to their high content of essential amino acids,high digestibility and biological value.Research in the past decades has shown that many of the whey protein fractions and peptides derived from them possess various biological activities.The technology developments in the last decade,such as those in the areas of protein fractionation technologies have made industrial scale production of fractionated whey protein products commercially feasible.The development in hydrolysis technology and the peptide analytical capability allowed production of whey protein hydrolysates with designed bioactivity and functionalities.People are continuously finding new biological activities of the various whey protein fractions or their hydrolysis products.Significant amount of research has also been carried out to study the clinical benefits of some of the whey protein fractions and whey protein derived peptides.Examples of these studies will be reviewed and discussed.

Research Progress of Anti-tumor Effects of Curcuma zedoaria and its Active Ingredients Through Immune Regulation Mechanism

The continuous increase in the incidence rate of various fatal malignant tumors in the recent years warrants an imperative search for medications or drugs with obvious anti-tumor eflects and reliable curative effects.Previous studies have found that Curcuma zedoaria and its active ingredients,such as turmeric oil,curcumol,and P-elemene,have obvious antitumor effects,and they do not have the adverse reactions and side effects seen in the anti-tumor drugs of Western medicine.Based on the review and inductive analysis of related literature,we summarize in the present article the results of some researchers who investigated the anti-tumor effects of Curcuma zedoaria and its active ingredients through the immune regulation mechanism.

Influence of alloy ingredients on mechanical properties of ternary boride hard alloy clad materials

Using Mo, B-Fe alloy and Fe powders as raw materials, and adding C, Cr and Ni ingredients, respectively, or C, Cr and Ni mixed powders, ternary boride hard alloy clad materials was prepared on Q235 steel substrate by means of in-situ reaction and vacuum liquid phase sintering technology. The influence of alloy ingredients on the mechanical properties of ternary boride hard alloy clad materials was investigated. The results indicate that a mixture of 0.8% C, 5% Cr and 2% Ni ingredients gives a ternary boride hard alloy clad material with optimal mechanical properties, such as high transverse rupture strength, high hardness and good wear resistance.

Developing new cancer nanomedicines by combination ancient traditional Chinese medicine effective ingredients and advanced nanotechnology

Various effective ingredients extracted from traditional Chinese medicine have achieved more and more attention in the field of cancer therapy due to their high bioactivity and good safety.These traditional Chinese medicine effective ingredients include paclitaxel,arsenical trioxide,camptothecin and so on.However,poor water solubility and low bioavailability of these traditional Chinese medicine ingredients severely hindered their clinical application.The rapid development of nanotechnology provided a new platform,new opportunities and new challenges for their effective delivery in vitro and in vivo.This minireview provides a concentrated summary of the most recent progresses made in this emerging field,highlighting the“combination ancient traditional Chinese medicine ingredients and advanced nanotechnology”strategy for favoring the construction of next generation cancer nanomedicines.

Sterilization of <i>Hibiscus rosa-sinensis</i>L. Vegetative Explants Sourced from Plants Grown in Open Environment and Influences of Organic Ingredients on <i>In Vitro</i>Direct Regeneration

This paper reports on the effects of organic ingredients in facilitating direct shoot regeneration from nodal explants of Hibiscus rosa-sinensis L. This paper also compares the sterilization conditions for 3 types of explants (node, internode, and shoot tip) harvested from an open field. The optimized sterilization conditions for the explants were 40% Clorox- 20 min exposure, 10% Clorox-15 min exposure, and 5% Clorox-40 min exposure for the node, internode and shoot tip, respectively. In the direct shoot regeneration using the nodal explants, we found MS medium containing 40 g/L sucrose, 0.3% (w/v) activated charcoal, and supplementations with myo-inositol, thiamine and nicotinic acid were suitable. The in vitro shoot survival rate was 30% with a mean leaf numbers of 2.68 produced, and a mean leaf length of 1.71 cm achieved after 5 weeks of culture on the modified medium.